Real-Time Single-Cell Measurement and Kinetic Modeling of Daunorubicin Uptake in Multidrug-Resistant Leukemia Cells...

Area of Science:

• Biomedical Engineering
• Cell Biology
• Pharmacology

Background:

• Multidrug resistance (MDR) is a key challenge in leukemia chemotherapy using anthracyclines like daunorubicin (DNR).
• Population-level drug uptake measurements mask individual cell variability, impacting therapeutic outcomes.
• Understanding single-cell drug kinetics is crucial for deciphering MDR mechanisms.

Purpose of the Study:

• To quantify real-time daunorubicin (DNR) uptake kinetics at the single-cell level in leukemia cells.
• To investigate the impact of P-glycoprotein (P-gp) mediated multidrug resistance (MDR) on DNR accumulation.
• To compare single-cell analysis with population-level measurements for assessing MDR heterogeneity.

Main Methods:

• Utilized a microfluidic biochip for long-term, real-time monitoring of DNR uptake in individual leukemia cells.
• Examined both drug-sensitive wild-type and multidrug-resistant (MDR) leukemia cells overexpressing P-gp.
• Employed a same-single-cell analysis (SASCA) approach to compare drug uptake before and after MDR inhibitor treatment.

Main Results:

• DNR uptake kinetics differed between sensitive (single process) and MDR (two processes) cells.
• Significant cell-to-cell heterogeneity in DNR uptake rates and retention was observed in both cell types.
• SASCA revealed enhanced DNR retention and uptake kinetics upon MDR inhibitor treatment, overcoming variability masking.

Conclusions:

• Real-time single-cell kinetic analysis uncovers functionally relevant heterogeneity in MDR leukemia.
• This approach provides deeper insights into MDR pathophysiology than population-averaged measurements.
• Understanding single-cell variability is essential for developing effective leukemia chemotherapy strategies.